Analysis of the NAT2 genotypes with real-time fluorescence quantitative polymerase chain reaction and its significance
10.3760/cma.j.issn.1009-9158.2011.09.017
- VernacularTitle:用实时荧光定量PCR检测NAT2基因型及其意义
- Author:
Ziyi LUO
;
Yumei WU
;
Jinfu PENG
;
Hongmei ZHANG
;
Bing LI
;
Saiyun LIU
- Publication Type:Journal Article
- Keywords:
Arylamine N-acetyhransferase;
Polymorphism,single nucleotide;
Polymerase chain reaction
- From:
Chinese Journal of Laboratory Medicine
2011;34(9):842-848
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore and analyze the distribution of NAT2 gene in Han population in Shenzhen andprovidethebasisfortreatmentof NAT2-relatedmetabolicdiseasesandcancer research. MethodThe study diluted the DNA sample(d0 ng/μl) as follows: 1 × 100, 1 × 101, 1 × 102, 1 ×103 , 1 × 104, to verify the sensitivity of detection of NAT2 gene polymorphism by real-time PCR. Mutation locus of 282,341,481, 590 and 857 of NAT2 in 554 normal samples were detected and genotyped by realtime PCR with Taqman probes. Forty-seven cases among 554 healthy controls were detected by DNA sequencing to verify the sensitivity, specificity and accuracy of this assay. ResultsThe lowest detection limit was 10-4 ng/μl. The frequencies of each allele of NAT2 were: *4 allele 64. 6％ (358/554), *5 allele 6. 3％ (35/554), *6 allele 25. 3％ ( 140/554), *7 allele 30. 0％ ( 166/554), * 11 allele 0. 6％ (3/554) The frequencies of main genotypes of NAT2 were as the follows : NAT2 *4/* 6 12.3％ ( 68/554 ), *6/* 78. 3％ (46/554), * 13/* 13 or * 12/* 12 or *4/* 4 7.6％ (42/554), *4/* 7 8. 1％ (45/554), * 7/* 1 312. 3％ (68/554), * 12 7.2％ (40/554), *6/* 13( * 12)5.6％ (31/554). The samples with 7 genotypes account for 89. 6％ (496/554) of the total cases.Rapid acetylation and Intermediate type accounting for 40. 5％ (224/554) and 46. 7％ (259/554) respectively were the main phenotypes of NAT2 in Han people in Shenzhen. In addition, the accuracy, specificity and sensitivity of the PCR were compared to direct DNA sequencing. The accuracy of 282, 341,481, 590 and 857 were 88.2％ (30/34), 87.5％ (7/8), 80. 0％(4/5), 100. 0％ ( 22/22 ) and 93.8％ ( 15/16 ) respectively. The specificities were 100. 0％ ( 13/13 ),94. 9％ ( 37/39), 100. 0％ ( 42/42 ) , 96.0％ ( 24/25 ) and 96. 8％ ( 30/31 ) respectively. The sensitivities were 91.5％ ( 43/47 ), 93.6％ ( 44/47 ), 97.6％ ( 46/47 ) 97.9％ ( 46/47 ) and 95.7％ ( 45/47 ),respectively. ConclusionsThe sensitivity, specificity and accuracy of real-time PCR established in this study is good. It can be widely used for clinical and scientific research. The major alleles of NAT2 of Han population in Shenzhen are * 4, *6, * 7. The most common slow acetylation genotype is *6/* 7. This study can provide a basis for NAT2-related metabolic diseases and cancer research.
